Abstract
S-palmitoylation is a pervasive post-translational modification required for the trafficking, compartmentalization and membrane tethering of many proteins. We demonstrate that the commercially available compound 17-octadecynoic acid (17-ODYA) can serve as a bioorthogonal, click chemistry probe for in situ labeling, identification and verification of palmitoylated proteins in human cells. We identified ∼125 predicted palmitoylated proteins, including G proteins, receptors and a family of uncharacterized hydrolases whose plasma membrane localization depends on palmitoylation.
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Acknowledgements
We thank E. Weerapana (The Scripps Research Institute) for providing reagents and assistance with click chemistry and mass spectrometry, G. Simon for assistance with data analysis, W. Kiossis for microscopy instruction and members of the Cravatt laboratory for helpful discussions. This research was funded by F32NS060559, CA087660 and the Skaggs Institute for Chemical Biology.
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B.R.M. performed experiments. B.R.M. and B.F.C. designed experiments, analyzed data and wrote the paper.
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Supplementary Figures 1–5, Supplementary Tables 1–2, Supplementary Methods (PDF 2340 kb)
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Martin, B., Cravatt, B. Large-scale profiling of protein palmitoylation in mammalian cells. Nat Methods 6, 135–138 (2009). https://doi.org/10.1038/nmeth.1293
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DOI: https://doi.org/10.1038/nmeth.1293
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